anti p stat1 tyr 701  (Cell Signaling Technology Inc)

Journal: Carcinogenesis

Article Title: Knockdown of ISOC1 inhibits the proliferation and migration and induces the apoptosis of colon cancer cells through the AKT/GSK-3β pathway

doi: 10.1093/carcin/bgz188

Figure Lengend Snippet: The effects of ISOC1 knockdown on the biological function of colon cancer cells are rescued by UCN01. Further verification of the mechanism of ISOC1 in colon cancer cells. The protein level of p-STAT Tyr 701 in colon cancer cell nuclei was increased by ISOC1 knockdown. ( A ) MTS assays. ( B ) Transwell migration assays. ( C ) Cell apoptosis assay. ( D ) Western blot assay. The protein level of p-GSK-3β Ser 9 was upregulated by overexpression of p-AKT Thr 308 . AKT Δ-Thr308 : p-AKT Thr 308 deletion plasmid. AKT P-Thr308 : p-AKT Thr 308 activation plasmid. ( E ) Western blot assay. The protein levels of c-Myc, MMP2 and MMP9 were significantly reduced by ISOC1 knockdown. ( F ) Confocal microscopy and western blot analysis of p-STAT1 Tyr 701 nuclear localization. Green signals represent cells infected with lentiviruses. DNA was visualized with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Western blot analysis showed that the protein levels of p-STAT1 Tyr 701 were increased in the shISOC1 group compared to the shCtrl group. The error bars represent the standard deviation. * P < 0.05, ** P < 0.01.

Article Snippet: The antibodies used for the western blotting included GAPDH (AP0063, Bioworld), ISOC1 (ab118245, Abcam), AKT1/2/3 (ET1609-51, Huabio, China), P-AKT Thr 308 (AF3262, Affinity), P-GSK-3β Ser 9 (AF2016, Affinity), P-STAT1 Tyr 701 (AF6300, Affinity), PARP1 (13371-1-AP, Proteintech), Caspase-3 (9662, CST), Caspase-8 (GB-11594, Servicebio, China), Caspase-9 (GB-11053-1, Servicebio, China), c-Myc (5605s, CST), MMP2 (4022s, CST) and MMP9 (3852s, CST).

Techniques: Knockdown, Migration, Apoptosis Assay, Western Blot, Over Expression, Plasmid Preparation, Activation Assay, Confocal Microscopy, Infection, Standard Deviation

Journal: bioRxiv

Article Title: Luteolin transforms the BMDM polarity to regulate the expression of inflammatory factors

doi: 10.1101/2020.06.30.181503

Figure Lengend Snippet: Effects of luteolin on the protein levels of p-STAT1/6 in polarized BMDM. The BMDM were primed with LPS+IFN-γ or IL-4, and then for addition of luteolin for 24 h. Total cell lysates were analyzed by immunoblotting for the indicated antibody, respectively. β-Actin was used as loading control. Representative immunoblots of p-STAT1 (A) and p-STAT6 (B); The relative protein levels of p-STAT1and p-STAT6 (D) by densitometric analysis. Data represented mean ± SD of three independent experiments. Different symbols indicate a significant difference according to ANOVA and Tukey test. * P <0.05 vs. Control group (without treatment); & P < 0.05 vs. LPS+IFN-γ-treated group.

Article Snippet: After 20 μg protein was subjected to SDS-PAGE and transferred to PVDF membrane (millipore, USA), the corresponding primary antibodies against p-STAT1-tyr 701 , p-STAT6-tyr 641 (Cell Signaling Technology, USA) and β-actin (Sigma, USA) were applied at 4°C overnight.

Techniques: Western Blot, Control

Journal: Molecular Metabolism

Article Title: Regulation of glucose uptake and inflammation markers by FOXO1 and FOXO3 in skeletal muscle

doi: 10.1016/j.molmet.2018.09.011

Figure Lengend Snippet: Effects of FOXOdn overexpression on inflammatory signaling pathways in tibialis anterior muscle . (A) Quantification of STAT1 protein content and phosphorylation after FOXO1dn or (B) FOXO3dn electroporation in tibialis anterior muscle. (C) Representative western blots of STAT1 protein content and phosphorylation after FOXO1dn or FOXO3dn overexpression. (D) Gene expression analysis of chemokine genes after FOXO1dn or FOXO3dn overexpression. (E) Gene expression of key immune cell markers after FOXO1dn or FOXO3dn overexpression. Data are mean ± SEM. n = 11 mice. * p < 0.05 and ** p < 0.01.

Article Snippet: The following antibodies used for immunoblot analysis were purchased from Cell Signaling Technology (Beverly, MA): Akt (#9272), p-Akt Thr 308 (#4056), p-Akt Ser 473 (#9271), GSK3β (#9315), p-GSK3β Ser 9 (#9323), GS (#3839), p-GS Ser 641 (#3891), mTOR (#2983), p-mTOR Ser 2448 (#5536), 4EBP1 (#9644), p-4EBP1 Thr 37/46 (#2855), p-p70S6K Thr 389 and Thr 421 /Ser 424 (#2708), p70S6K (#9205), p-STAT1 Tyr 701 (#9171), STAT1 (#9172).

Techniques: Over Expression, Electroporation, Western Blot, Expressing